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Fluoro SSAO™
Fluorescent Semicarbazide-Sensitive Amine Oxidase Detection Kit

Key Benefits:
  • Non Radioactive
  • Can monitor multiple time points to follow kinetics.
  • Monitors Enzymatic Activity
  • One-step, no wash assay.
  • Adaptable for High Throughput format
  • Enzyme Positive Control included in Kit
  • Applications - Fluorescence Plate Reader
Assay Principle
Semicarbazide-sensitive amine oxidase (SSAO) is a common name for a widely distributed enzyme in nature. In man this enzyme is present in the vascular system and circulates in plasma. SSAO differ from the monoamine oxidases A and B in substrate and inhibitor patterns. These enzymes have been widely studied and their tissue distribution, molecular properties, substrate specificities and inhibitor sensitivities are extensively reviewed (2,3).

SSAO exists in two forms: tissue bound and soluble (plasma SSAO). Tissue bound SSAO acitvity is associated with blood vessels, mainly in smooth muscle layers, however it is also associated with spleen, placenta, bone marrow, kidney, sclera, retina, endothelial cells, adipocytes, chondrocytes and fibroblasts. (4,5). Evidence suggests that Plasma SSAO originates from the cleavage of membrane-bound form. The possible sources of plasma SSAO are still unclear, but it has been suggested that it may be derived from liver, retina, placenta and bone tissue (6,7,8).

SSAO’s functional role has been suggested to be involved in: apoptosis, atherogenesis, cell adhesion, leucocyte trafficking, glucose transport and local production of hydrogen peroxide. Reports of elevated levels of SSAO have been reported in congestive heart failure, diabetes mellitus, alzheimer’s disease and various other inflammatory diseases (1).

Furthermore byproducts of SSAO deamination, such as formaldehyde and methylglyoxal, have been proposed to be involved in pathogenesis of cancer, aging and atherosclerosis.

The Fluoro SSAO detection kit utilizes a non- fluorescent detection reagent, to detect H202 released from the conversion of Benzylamine to Benzaldehyde via SSAO. Furthermore H202 oxidizes this detection reagent in a 1:1 stoichiometry to produce a fluorescent product resorufin. This oxidation is catalyzed by Peroxidase.

Reaction:
Benzylamine + O2+ H2O + SSAO Benzaldehyde + NH3 + H202

H202 + Detection reagent (non-fluorescent)+Peroxidase Resorufin (fluorescent)

References
  1. Yayin Tahiri Kasim et.al., Turk J Biochem; 29 (3);247-254 (2004); Semicarbazide-Sensitive Amine Oxidase: Biochemical and Physiological Properties
  2. B.A.Callingham,A.Holt,J.Elliott.J.Neural Transm.(Suppl.)32 ,279 –290 (1990).
  3. B.A.Callingham and M.A.Barrand.J.Neural Transm.(Suppl.)23 ,37 –54 (1987).
  4. G.A.Lyles.J.Neural Transm.(Suppl.)41 ,387 –396 (1994).
  5. K.Magyar, Z.Mészáros and P.Mátyus . Semicarbazide-sensitive amine oxidase.Its physiological significance Pure Appl.Chem.,Vol.73,No.9,pp.1393 –1400,2001.
  6. Boomsma F.,van Dijk J.,Bhaggoe U.M.,Bouhuizen A.M.B.,van den Meiracker A.H.(2000)Variation in semicarbazide-sensitive amine oxidase activity in plasma and tissues of mammals.Comp.Biochem.
  7. Ekblom J.,Gronval J.L.,Garpenstrand H.,Nillson S.,Oreland L.(2000)Is semicarbazide-sensitive amine oxidase in blood plasma partly derived from the skeleton?.Neurobiology,8,129-135.
  8. McGuirl M.A., Dooley D.M.(1999)Copper-containing oxidases. Curr.Opin.Chem.Biol.,3,138-144.
  9. Methods Enzymology 142, 617 (1997).
  10. Holt A. ; Sharman D.F. ; Baker G.B. ; Palcic M.M. A Continuous Spectrophotometric Assay for Monoamine Oxidase and Related Enzymes in Tissue Homogenates Analytical Biochemistry, January 1997, vol. 244, no. 2, pp. 384-392(9).
  11. Biochem Pharmacol 18, 1447 (1969).
Kit contents (for 500 assays)
Description
Part#
Storage after opening Kit or Reconstitution of Reagents.
1 Bottle: 5X Reaction Buffer pH 7.4
3019 4-80C
1 vial: Detection reagent for 500 Assays 4008
Aliquot in single use vials: Below –200C
1 vial: Horseradish Peroxidase 6005 4-80C
1 vial: SSAO substrate Benzylamine
7001 Aliquot in vials: Below –200C
1 vial: SSAO Enzyme 6006 Aliquot in single use vials: Below –200C
1 vial Pargyline: Monoamine Oxidase B inhibitor (9-11) 7003 Aliquot in vials: Below –200C
1 vial Semicarbazide: Semicarbazide-sensitive amine oxidase inhibitor. 7004 Aliquot in single use vials: Below –200C
The following kits are available:
Catalog #
Size
Price (US$)
SSAO 100-3
500 Tests $295
* Please call 888 7 ASSAYS (888-727-7297) or email info@celltechnology.com for volume pricing
 
 Product Data Sheet
 Protocol
 MSDS